RhoC expression and head and neck cancer metastasis.
RhoC expression and head and neck cancer metastasis.
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DOI：
10.1158/1541-7786.mcr-08-0512
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发表时间：
2009-11
期刊：
影响因子：
--
通讯作者：
Teknos TN
中科院分区：
文献类型：
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作者：
Islam M;Lin G;Brenner JC;Pan Q;Merajver SD;Hou Y;Kumar P;Teknos TN
RhoC蛋白是侵袭性乳腺癌和黑色素瘤转移的已知标志物，在某些头颈癌中也被发现过表达，因此我们研究了RhoC表达与头颈鳞状细胞癌转移行为的相关性。利用慢病毒小发夹RNA （shRNA）进行转导，并用绿色荧光蛋白（GFP）跟踪，实现了RhoC表达的选择性抑制，达到70-80%的RhoC抑制。在RhoC敲低稳定克隆的荧光显微镜下，大多数细胞显示强烈的绿色荧光，表明转导效率高。重要的是，qRT-PCR显示Ras超家族其他成员的mRNA表达水平没有显著降低。与对照转导细胞系相比，RhoC缺失细胞系的细胞运动性和侵袭性明显减弱。植入RhoC敲低细胞的SCID小鼠肺组织的苏木精和伊红染色显示肺转移和血管炎症明显减少。与shRNA打乱序列控制系相比，植入RhoC缺失细胞系的小鼠培养的肺组织细胞生长明显下降。CD31表达的显微镜研究显示，与亲代和shrna混乱的对照组相比，原发肿瘤微血管密度在数量和质量上存在实质性差异。这项研究首次明确了RhoC在头颈部转移中的作用。这些发现表明，RhoC作为一种新的潜在治疗靶点，其稳健性值得进一步研究。
RhoC protein, a known marker of metastases in aggressive breast cancers and melanoma, has also been found to be over-expressed in certain head and neck cancers, thus we investigated the correlation between RhoC expression and the metastatic behavior of head and neck squamous cell carcinoma. Selective inhibition of RhoC expression was achieved using lentiviral small hairpin RNA (shRNA) transduced and tracked with green fluorescent protein (GFP) to achieve 70-80% RhoC inhibition. Fluorescence microscopy of the RhoC knockdown stable clones showed strong green fluorescence in the majority of cells, signifying a high efficiency of transduction. Importantly, qRT-PCR showed no significant decrease in the mRNA expression levels of other members of the Ras superfamily. Cell motility and invasion were markedly diminished in RhoC depleted cell lines as compared to control transduced lines. Hematoxylin and eosin staining of lung tissue obtained from SCID mice which had been implanted with RhoC knockdown cells showed marked decrease in lung metastasis and inflammation of the blood vessels. The cultured lung tissue showed a significant decrease in cell growth in mice implanted with RhoC depleted cell lines as compared to shRNA scrambled sequence control lines. Microscopic studies of CD31 expression revealed substantial quantitative and qualitative differences in the primary tumor microvessel density as compared to parental and shRNA-scrambled controls. This study is the first of its kind to establish the involvement of RhoC specifically in head and neck metastasis. These findings suggest that RhoC warrants further investigation to delineate its robustness as a novel potentially therapeutic target.